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CHARACTERIZATION OF NUTRIENT DEFICIENCY

IN

Hancornia speciosa

Gomes SEEDLINGS BY OMITTING

MICRONUTRIENTS FROM THE NUTRIENT SOLUTION

1

LAYARA ALEXANDRE BESSA2, FABIANO GUIMARÃES SILVA3,

MARIALVA ALVARENGA MOREIRA4, JOÃO PAULO RIBEIRO TEODORO5,

FREDERICO ANTÔNIO LOUREIRO SOARES3

ABSTRACT - Hancornia speciosa Gomes (Mangaba tree) is a fruit tree belonging to the Apocynaceae

family and is native to Brazil. The production of seedlings of this species is limited by a lack of technical and nutritional expertise. To address this deficiency, this study aimed to characterize the visual symptoms of micronutrient deficiency and to assess growth and leaf nutrient accumulation in H. speciosa seedlings supplied

with nutrient solutions that lack individual micronutrients. H. speciosa plants were grown in nutrient solution

in a greenhouse according to a randomized block design, with four replicates. The treatments consisted of a group receiving complete nutrient solution and groups treated with a nutrient solution lacking one of the following micronutrients: boron (B), copper (Cu), iron (Fe), manganese (Mn), zinc (Zn), and molybdenum (Mo). The visual symptoms of nutrient deficiency were generally easy to characterize. Dry matter production was affected by the omission of micronutrients, and the treatment lacking Fe most limited the stem length,

stem diameter, root length, and number of leaves in H. speciosa seedlings as well as the dry weight of leaves, the total dry weight, and the relative growth in H. speciosa plants. The micronutrient contents of H. speciosa leaves from plants receiving the complete nutrient solution treatment were, in decreasing order,

Fe>Mn>Cu>Zn>B.

Index terms: Hancornia speciosa Gomes, mineral nutrition, symptoms, growth.

CARACTERIZAÇÃO DE DEFICIÊNCIA NUTRICIONAL DE

MUDAS DE MANGABEIRA

(Hancornia speciosa

Gomes) COM

OMISSÃO DE MICRONUTRIENTES EM SOLUÇÃO NUTRITIVA

RESUMO - A mangabeira (Hancornia speciosa Gomes) é uma frutífera pertencente à família Apocynaceae,

originária do Brasil. A produção de mudas desta espécie em grande escala ainda é limitada pela escassez de conhecimentos técnicos e nutricionais. Para obter maiores esclarecimentos sobre o assunto, este trabalho objetivou caracterizar os sintomas visuais de deficiência de micronutrientes, avaliar o crescimento e o acúmulo de nutrientes foliar de mudas de mangabeira, submetidas a diferentes soluções nutritivas com suspensão de

micronutrientes. Plantas de mangabeira foram cultivadas em solução nutritiva, dentro de casa de vegetação,

no delineamento de blocos casualizados, com quatro repetições. Os tratamentos foram constituídos de solução nutritiva completa e omissão individual de boro, cobre, ferro, manganês, zinco e molibdênio. Os sintomas visuais de deficiências de nutrientes mostraram-se, de maneira geral, facilmente caracterizáveis. A produção de matéria seca foi afetada pela omissão de micronutrientes, e o tratamento que mais limitou o

comprimento do caule, diâmetro do caule, comprimento da raiz, número de folhas de mudas de mangaba e massa seca das folhas, massa seca total e o crescimento relativo da mangabeira, foi a omissão de ferro. Os conteúdos de micronutrientes das plantas do tratamento completo obedeceram à seguinte ordem decrescente

de micronutrientes Fe>Mn>Cu>Zn>B em folhas de mangabeira.

Termos para indexação: Hancornia speciosa Gomes, nutrição mineral, sintomas, crescimento.

1(Trabalho 097-12). Recebido em: 27-02-2012. Aceito para publicação: 28-03-2013.

2Bióloga, mestra em Ciência Agrárias, Instituto Federal Goiano - Câmpus Rio Verde, caixa postal 66, CEP: 75901-970 - Rio Verde –

GO. E-mail: layara.ifmorrinhos@hotmail.com (autor para correspondência)

3Professor do Instituto Federal Goiano - Câmpus Rio Verde – GO. E-mails: fabianoifgoiano@gmail.com; fredalsoares@hotmail.com 4Bolsista PNPD CAPES, Instituto Federal Goiano - Câmpus Rio Verde – GO

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INTRODUCTION

The Hancornia speciosa Gomes tree

(Mangaba tree), from the Apocynaceae family, is

native to Brazil and has a wide geographical range, occurring in ecosystems such as the Amazon, Atlantic

Forest, and Cerrado (ROSA et al. 2005). This plant

produces edible fruits that can be consumed fresh and used to produce juices, candy, and ice cream

(SOARES et al. 2011).

In Brazil, the production of H. speciosa fruit

in 2010 was approximately 722 tons (IBGE, 2010). With the exception of the state of Minas Gerais, there

are only harvest records for this fruit in the Northeast region, and the states of Sergipe, Bahia, and Paraíba are currently the largest producers, with production totals of 401, 142, and 99 tons, respectively. The state of Minas Gerais produced only one ton of H. speciosa fruit, (IBGE, 2010).

Seedling quality is an essential factor

for homogeneity, rapid development, and early

production, thus ensuring the expansion of this crop in Brazil. To obtain high-quality seedlings, it is necessary to adequately meet their nutrient demands (NATALE et al. 2006). Currently, the management

of H. speciosa seedling nutrition is performed

empirically by the nursery workers, in view of the lack of expertise in the nutritional aspects of this crop, which results from a lack of scientific studies. The exploitation of a native species depends on technical expertise that is generally scarce; therefore, there is a need for research to make their exploitation and market insertion (ROSA et al. 2005).

To expand the production of the fruits,

productive factors, such as the nutrient supply, need

to be controlled because nutrient levels at specific

stages of plant development affect productivity.

(TAIZ; ZEIGER, 2009).

The nutrients perform specific functions in the life of the plant; thus, when plants are subjected

to nutrient-omission treatments, they show visual

symptoms of deficiency when the level received by the plant is below what is considered adequate. In general, each nutrient has specific functions in the

plants, and the different elements are associated

with different symptoms of deficiency or toxicity.

A deficiency in one specific nutrient does not necessarily affect the same metabolic process in all

plant species (TAIZ; ZEIGER, 2009).

The probability of a micronutrient deficiency is lower than that of a macronutrient deficiency. Zn (zinc) and B (boron) are the most commonly

deficient micronutrients in Brazilian soils, and

Fe deficiency may occur due to excessive liming

or due to competitive inhibition by metals such

as Mn (manganese), Cu (copper) and Zn (zinc) (MALAVOLTA, 2006 ).

Leaf analysis of fruit-bearing plants has been used to detect plant responses to several

types of management, thus enabling more efficient

interpretation of the relationships between nutrients in

the plant (SILVA; CARVALHO, 2005). Furthermore, it allows for the identification and correction of deficiencies (SARCINELLI et al. 2004), thus making

better use of fertilizers, leading to savings of this

input and, consequently, achieving better product quality.

Considering that H. speciosa is in the process

of being domesticated, it is necessary to know the best planting techniques and nutrient requirements

for this tree. Growing plants in nutrient solution has been an important tool in advancing plant nutrition research because the composition of the solution is

controlled and the complexity of soil composition

is eliminated. Furthermore, studies with plants in a controlled environment suggest that the effects of different nutrients on plant growth can be more easily

studied than in the field (TAIZ; ZEIGER, 2009).

Therefore, this study aimed to characterize the visual

symptoms of micronutrient deficiency and to evaluate

the growth and the leaf nutrient accumulation of

H. speciosa seedlings exposed to different nutrient

solutions with micronutrient omissions.

MATERIALS AND METHODS

The experiment was conducted in the greenhouse of the Laboratório de Cultura de Tecidos Vegetais at the Instituto Federal Goiano – Campus Rio Verde, from January to June 2011. The upper,

front and side portions of the structure were covered

with a 0.1mm thick transparent polyethylene film.

The H. speciosa plants used were taken from the

Gameleira farm, which is located in the municipality of Montes Claros, in the state of Goiás, at a height of 592 m above sea level and with the geographical

coordinates of 16° 06’ 20’’ S and 51° 17’ 11” W. The seeds were manually extracted, using a sieve to help remove the pulp, and subsequently

washed in running water. They were then planted

in 72 cell expanded polystyrene trays filled with vermiculite and carbonized rice husks at a ratio of

1:1. Thirty days after planting, seedlings with 3-4 full leaves were selected, roots were rinsed with

deionized water and transferred to 8L pots containing Hoagland and Arnon (1950) nutrient solution at 50%

of the original ionic strength, for a period of 30 days.

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nutrient-omission treatment. The control group received the complete solution, and other groups were treated

with a nutrient solution lacking one of the following micronutrients: B, Cu, Fe, Mn, Zn, and Mo, for a total of 28 plots. Each experimental plot consisted of an 8-L pot of nutrient solution containing two plants. The treatments were arranged in randomized block

design, with four replicates.

During the experimental period, the plants

were grown under an average irradiance of 200 µmol×m-2s-1. The nutrient solution was kept under

constant aeration using compressed air, and the

pH was adjusted daily to 5.5 ± 0.5 by adding HCl or NaOH as needed. The nutrient solution was

replaced when the initial electrical conductivity value

decreased by 30%.

Every day, the visual symptoms of

nutrient deficiency were assessed, recorded, and

photographed.

Thirty days after transplantation (DAT),

the plant growth was assessed for each treatment

solution by examining the stem length, using a ruler graduated in centimeters (cm) from the root collar

to the terminal bud, and the diameter was measured

using a digital caliper graduated in millimeters; the

number of leaves was obtained by counting, and the length of the longest root was measured using a ruler graduated in cm. These measurements were

taken every 15 days until the plants were harvested, as the deficiencies caused by the nutrient omission

became more pronounced. The cultivation cycle

ended at 120 DAT for the treatments in which B, Zn,

and Mo were omitted and for the treatment with the complete nutrient solution, and the cycle ended at

60 and 105 DAT for the treatments in which Fe and

Cu were omitted, respectively, because the seedlings did not survive past these times. The plants were

harvested and taken to the laboratory, where they

were sectioned into leaves, stem and roots. These organs were weighed and then placed in an oven at

65 °C until they reached a constant mass, to obtain

their dry matter weights.

After drying, the leaves were crushed in a

Wiley mill equipped with a 20-mesh sieve and sent to

the leaf analysis laboratory to determine their B, Cu,

Mn, Zn and Fe contents, following the methodology proposed by Malavolta et al. (1997).

The amounts of each nutrient in the leaves were calculated by multiplying the dry weight of the leaves by the content of each nutrient in the leaves.

The shoot-to-root ratio (S/R) was calculated

by adding the dry matter weights of the stem and leaves and then dividing by the dry matter weight

of the root. The relative growth (RG) corresponds

to one hundred times the ratio of the total dry matter from the nutrient-omission treatments to the total dry matter from the complete nutrient treatment

(BENINCASA, 2003)

The data obtained were subjected to an analysis of variance, and the means were compared

by Tukey’s test at 5% probability. The observed

values for the stem dry matter, root dry matter, leaf dry matter, total dry matter, relative growth, and shoot-to-root ratio needed to be transformed into

the root of x+1 to obtain a normal distribution and

homogeneity of variances.

RESULTS AND DISCUSSION

Visual symptoms of nutrient deficiency

The visual symptoms of B deficiency were

initially observed at 40 DAT. The younger leaves showed chlorosis followed by necrosis, which began in the leaves and moved towards the terminal buds,

with a loss of apical dominance (Figure 1). There

was also premature leaf abscission and reduction in

the length of the stem and roots. Neves et al. (2004)

observed that the symptoms of B deficiency in

Spondias tuberosa Arr plants (Umbu) were clearly

identifiable 20 days after starting treatment. The

deficiency was observed in the younger leaves, which showed mild internerval chlorosis that

subsequently expanded from the centers to the edges

of the leaves. Similar symptoms were observed by

Viégas et al. (2004) in camu-camu trees (Myrciaria dubia (H.B.K.) McVaugh) at 50 days after starting

treatment. Young leaves were twisted, atrophied, small, and thick. As the symptoms intensified, the

shoot apical meristem died.

For Cu, the visual symptoms of deficiency began at 35 DAT. Young leaves showed darkening with necrotic spots that began at the apex and

advanced towards the leaf base along the edges

(Figure 1). Wrinkling of the leaves was also observed.

The visual symptoms of Mn deficiency

were initially observed at 27 DAT. In older leaves, internerval chlorosis was observed under a thick

grid of veins that remained green. As the symptoms developed necrotic spots emerged and the leaves

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Zn omission resulted in the first visual

symptoms of deficiency at 30 DAT. Initially,

new leaves showed chlorosis. As the deficiency intensified, the plants showed smaller, wrinkled

leaves, a shortening of internodes, and

well-developed roots (Figure 1). Neves et al. (2004)

observed similar symptoms in Spondias tuberosa

Arr plants, which produced small and narrow leaves,

with chlorotic areas between the veins. Zn deficiency

compromises important physiological events and,

consequently, the growth and development of the plant (ZABINI et al. 2007).

The Fe omission treatment was the first

to show deficiency symptoms at 25 DAT. Young

leaves showed chlorosis, which started at the base

of the petiole and progressed towards the apex of the

leaves. As the symptoms evolved, the leaves became

smaller and completely whitened (Figure 1). The

roots showed limited growth and light coloration.

Neves et al. (2004) observed that the symptoms of Fe deficiency in Spondias tuberosa Arr plants started at 30 days, with changes in the color of younger leaves and a purple hue in the stem, sharply contrasting with the yellow remainder of the lamina.

The consequences of an insufficient Fe supply in fruit species, such as peach, pear, citrus, and kiwi, include symptoms like chlorosis, with consequent limitations on productivity, fruit quality, and plant vigor (CESCO et al. 2006).

For Mo, the visual symptoms of deficiency were initially observed at 60 DAT. There was

chlorosis of older leaves evolving to necrosis, with

early leaf abscission (Figure 1). Mo deficiency can

decrease nitrogenase enzyme synthesis, promoting

a reduction in biological nitrogen fixation and, therefore, reducing productivity (MORAES et al. 2008).

Plant growth and Dry matter production

The stem length of plants grown in a

nutrient-lacking solution, with the exception of Mo omission,

was reduced relative to treatment with a complete nutrient solution. Over the production cycle of H. speciosa seedlings, plants grown in a solution lacking

Fe showed the greatest reduction in stem length at

60 days (6.33 cm) relative to plants grown in the complete solution (7.10 cm) (Figure 2).

The first visual symptoms of deficiency in

H. speciosa plants occurred in the Fe-omission

treatment, resulting in the harvest of plants at 60 DAT. The micronutrient deficiencies negatively

affected plant growth, reducing stem length during the production cycle of H. speciosa seedlings, in the

following order: Fe>Cu>Zn>B>Mn>Mo.

The micronutrient omission treatments restricted stem diameter development in H. speciosa

plants when compared to complete nutrient treatment

(Figure 3). Fe omission caused the greatest restriction

in stem diameter development for H. speciosa

seedlings; stem diameters averaged 0.77 mm at 60 days, compared to 1.59 mm for seedlings

receiving the complete treatment. Neves et al.

(2004) observed a similar behavior in Spondias tuberosa Arr seedlings. Micronutrient deficiency

in H. speciosa seedlings negatively affected the development of stem diameter, in the decreasing

order Fe>Mo>Mn>Zn>B>Cu.

With the exception of plants grown under

the Mn-omission treatment, the other treatments restricted the number of H. speciosa leaves produced compared with the complete nutrient solution

(Figure 4). The solution lacking Fe was the most

limiting in terms of the number of leaves, with

plants grown in this solution having 5 leaves at 60 days, compared with 7 leaves for plants grown in the complete solution. There was a 28.57% reduction in

the number of leaves for the Fe-omission treatment compared to the complete solution. Neves et al.

(2004) also observed that Fe was the most limiting

element for leaf number in Spondias tuberosa Arr. Micronutrient deficiency in H.speciosa

seedlings negatively affected the number of l ea v es p r o d u c ed , i n t h e d e c r e a s in g o r d e r

Fe>Mo>B>Cu>Zn>Mn.

Root length was restricted in H. speciosa

plants under micronutrient omission treatment when compared to plants treated with the complete

nutrient solution (Figure 5). The Fe-deficient solution

caused the greatest restriction in root growth, with

roots averaging 9 cm in length at 60 days compared

with 12.34 cm for plants treated with the complete

nutrient solution. There was a 27.06% reduction

in root length for the Fe-omission treatment when compared with the values shown by the plants treated with the complete solution. Fruit species

that are Fe deficient show developmental, vigor, and productivity limitations (CESCO et al. 2006).

The micronutrient deficiency in H. speciosa

seedlings negatively affected root growth, according

to the following order Fe>Mo>Zn>B>Cu>Mn.

Fe omission in the nutrient solution was the most limiting treatment for the production of leaf dry matter, total dry matter, and relative growth, differing from the complete treatment in all of these

variables. There was a reduction of 72.54%, 68.36%, and 67.44%, respectively, when the Fe-omission

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treatment (Table 1). Neves et al. (2004) observed

that Fe omission was the most limiting treatment for the accumulation of dry weight of leaves, total dry

weight, and relative growth, with reductions of 80%, 75%, and 74.93%, respectively, compared with the

complete nutrient treatment for Spondias tuberosa

Arr plants.

Leaf nutrient content

The micronutrient contents of plants under the complete nutrient treatment and those from the nutrient-omission treatments were: B = 2.0 and 1.0 mg plant-1; Cu = 8.0 and 0.04 mg plant-1; Mn = 20.0

and 0.6 mg plant-1; Zn = 7.0 and 3.0 mg plant-1; and

Fe = 41.0 and 2.00 mg plant-1, respectively (Table

2). The relative micronutrient contents of plants

treated with the complete solution followed the order

Fe>Mn>Cu>Zn>B.

All of the micronutrient contents in plants grown under micronutrient-omission treatments were lower than those found for the complete solution

treatment, except for the B (Table 2). A deficiency

in one nutrient may cause an imbalance in other

nutrients and consequently lead to morphological

changes that are translated into the characteristic

symptoms of deficiency of each nutrient (LANGE

et al. 2005).

Although Malavolta et al. (1997) considered competitive inhibition between Cu, Zn, and Fe

common, it was not observed in this study. The ability

of Cu to displace ions, especially Fe and Zn, from exchange sites is the main cause of this inhibition.

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FIGURE 2 - Stem length (cm) of Hancornia speciosa Gomes plants, as a function of days after transplanting for the solution with the omission of micronutrients and nutrient solution.

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FIGURE 4 - Number of leaves in Hancornia speciosa Gomes plants, as a function of days after transplanting for the solution with the omission of micronutrients and nutrient solution.

FIGURE 5 - Root length (cm) of Hancornia speciosa Gomes plants, as a function of days after transplanting for the solution with the omission of micronutrients and nutrient solution.

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CONCLUSIONS

1-The visual symptoms of nutrient deficiencies

were generally easy to characterize.

2- Dry matter production was affected by micronutrient omission, and Fe omission was the treatment that most strongly affected the characteristic of plant.

3- The micronutrient contents of plants treated with the complete nutrient solution accumulated micronutrients in the following order:

Fe>Mn>Cu>Zn>B.

ACKNOWLEDGEMENTS

We thank the Brazilian Federal Agency for

the Support and Evaluation of Graduate Education

(Coordenação de Aperfeiçoamento de Pessoal de

Nível Superior — CAPES) for a post-doctoral scholarship and the Goias Research Foundation

(Fundação de Amparo à Pesquisa do Estado de

TABLE 1- Leaf dry matter (LDM), stem dry matter (SDM), root dry matter (RDM), total dry matter (TDM),

relative growth (RG), and shoot-to-root ratio (S/R) in Hancornia speciosa Gomes seedlings

grown in complete nutrient solution or in solutions lacking individual micronutrients. Rio Verde. Federal Institute of Goias. 2011.

TREATMENTS LDM(g) SDM(g) RDM(g) TDM(g) RG

% S/R

COMPLETE 0.511 a2 0.22 a 0.25 a 0.98 a 100 a 3.12 a

- B 0.28 ab 0.08 a 0.17 a 0.54 ab 57.65 ab 2.79 a

- Cu 0.19 ab 0.05 a 0.08 a 0.33 b 34.70 b 2.69 a

- Mn 0.28 ab 0.09 a 0.12 a 0.51 b 54.22 ab 3.04 a

- Zn 0.32 ab 0.08 a 0.23 a 0.64 ab 73.73 ab 2.07 a

- Fe 0.14 b 0.05 a 0.12 a 0.31 b 32.56 b 1.63 a

- Mo 0.19 ab 0.07 a 0.11 a 0.37 b 39.71 b 3.00 a

CV (%)3 5.41 3.30 3.32 6.22 18.20 20.06

1Means shown with original values; 2Means followed by the same letter in the same column are not significantly different according to

Tukey’s test at 5% probability; 3Coefficient of variation obtained with the data transformed into the root of x+1.

TABLE 2- Leaf content (mg plant-1) in Hancornia speciosa Gomes seedlings grown in complete nutrient

solution or in solutions lacking individual micronutrients. Rio Verde. Federal Institute of

Goias. 2011.

CONTENT (mg plant-1)

TREATMENTS B Cu Mn Zn Fe

COMPLETE 2.01a2 8.00 a 20.0 a 7.0 a 41.0 a - B 1.0 a 0.10 b 0.80 b 1.0 b 14.0 b

- Cu 1.0 a 0.04 b 2.00 b 0.5 b 10.0 b

- Mn 1.0 a 0.04 b 0.60 b 1.0 b 16.0 b

- Zn 1.0 a 2.00 b 10.0 a 3.0 b 24.0 ab

- Fe 0.6 a 0.01 b 0.20 b 0.1 b 2.00 b

CV (%)3 0.04 0.08 0.25 0.08 0.51

1Means shown with original values; 2Means followed by the same letter in the same column are not significantly different according to

Tukey’s test at 5% probability; 3Coefficient of variation obtained with the data transformed into the root of x+1.

Goiás — FAPEG) for financial support. We thank

the owner of Gameleira Farm for donating the plant material used in this study.

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Imagem

FIGURE 1 -  Nutrient deficiency symptoms in Hancornia speciosa Gomes seedlings grown in a complete  nutrient solution and in solutions lacking individual micronutrients (B, Cu, Mn, Zn, Fe and Mo)
FIGURE  3 - Stem  diameter  (mm)  of  Hancornia speciosa Gomes plants, as a function of days after  transplanting for the solution with the omission of micronutrients and nutrient solution.
FIGURE 5 -  Root length (cm) of  Hancornia speciosa Gomes plants, as a function of days after transplanting  for the solution with the omission of micronutrients and nutrient solution.
TABLE 1- Leaf dry matter (LDM), stem dry matter (SDM), root dry matter (RDM), total dry matter (TDM),  relative growth (RG), and shoot-to-root ratio (S/R) in Hancornia speciosa Gomes seedlings  grown in complete nutrient solution or in solutions lacking in

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Figure 2 - Percentage of spores in different phases of Bryum argenteum after fourteen days in distilled water and nutrient solution at 12 h photoperiod and 25º C.. Symbols

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Figure 12- Frame 0: Interactions between NMT and X25, the ligand molecule is indicated in purple on the active site of the enzyme, the dark red shapes indicate

evaluate the effect of the flow rate of the nutrient solution and the CE concentration (measured through the ionic strength of the solution) over the accumulation of fresh mass

Os exemplos seguintes são precisamente sequências que ilustram diferenças típicas entre a oralidade e a escrita, ou seja, entre o texto de partida e de chegada,

The visualization tool allows close access to the Markov chain library and easily manage the data modeling. Also, once the library finalizes the model, the tool displays it in

Estruturou-se assim, então este relatório de investigação que está composto por cinco capítulos, sendo o primeiro dedicado ao referencial teórico onde destacamos as